Program Director/Principal Investigator (Last, First, Middle): Vamum, Susan Marie PROJECT SUMMARY (See instructions): There are a number of toxins, includingbotulinumneurotoxin, ricin, shiga toxin and Staphylococcus enterotoxin B (SEB),that have either been used as bioterrorism agents or are considered a bioterrorism threat because of their extreme toxicity or ease of availibility. Furthermore, several of these toxins (botulinum neurotoxin, ricin) work at low concentrations, requiringhighlysensitive assays for detection. Additionally, it is important to distuiguish between active and inactive toxin because of the possibilitythat genetically engineered toxins, consisting of the enzymatic portion of the toxin and a bindingdomain of another protein, can be used as the bioweapon agent. Consequently,our long-term goal is to develop diagnostic assays for toxin detection and differentiation that address these dual requirements of high sensitivity and determination of toxin activity. Our objective in this proposal is to develop a platform for toxin detection based upon a combination of two analytical approaches: 1) molecular recognition based upon ELISA microarrays and 2) enzymaticactivity assays for toxin activity. We propose that this combined platform will provide a highly sensitive and specfic assay for the detection of these toxins. The specific aims are: 1). Developan ELISA microarray for the sensitive and quantitative detection of botulinum neurotoxins (serotypes A,B,C,D,E,and F), ricin toxin, shiga toxin and Staphylococcal enterotoxin B. 2). Functionalactivity assays specific for botulinumneurotoxins, ricin, and shiga toxin will be developed. 3). A single platform, comprised of the ELISA microarray and the activity assay will be developed, allowing for the sensitive and specificdetection and the determination of functional activity of,botulinumneurotoxins,ricin, Shiga toxin and Staphylococcal enterotoxin B in clinical samples. The combined assay platformwill be used to analyze clinical samples, includingsera, nasal swabs and stool samples, and for the determination of the sensitivity and specificityof the assays. RELEVANCE (See instructions): In the event of a bioweapon attack, it will be critical to rapidly determine which bioweapon agent is responsible for the attack in order to give the appropiate medical care. Our objective in this proposal is to develop a sensitive and specific assay for the detection of the following toxins: botulinum neurotoxin, ricin toxin, shiga toxin and Staphylococcus enterotoxin B. PROJECT/